抗纤软肝颗粒对大鼠肝星状细胞周期的影响

Effects of Kangxian Ruangan granule on hepatic stellate cell cycle of rat

目的:观察中药抗纤软肝颗粒对大鼠肝星状细胞(HSC)周期的影响。方法:SD雄性大鼠以抗纤软肝颗粒灌胃制备药物血清,HSC-T6细胞株复苏后置于培养箱内于37℃、5%CO2条件下培养,分为药物组(抗纤软肝颗粒用含5%FCS的DMEM培养基倍比稀释成5mg/ml、2.5mg/ml、1.25mg/ml 3个浓度),含药血清组(加5%、10%、20%3个不同浓度的抗纤软肝颗粒药物血清),空白对照组(加含5%FCS的DMEM培养基)及正常血清组(加正常大鼠血清)。MTT比色法测细胞增殖的抑制率,流式细胞仪测细胞周期。结果:药物组及含药血清组均能抑制活化HSC的增殖,各药物组及含药血清组均明显优于其对照组(P<0.01,P<0.05);细胞G0/G1期DNA含量的比较,10%及20%含药血清组明显高于正常血清组(P<0.01,P<0.05),药物组均明显高于其对照组(P<0.01,P<0.05)。结论:抗纤软肝颗粒能阻滞HSC从G1期向GS期的进行(即阻滞HSC通过G1/S关卡),造成G1期细胞堆积,阻滞DNA合成和复制,起到抑制HSC增殖的作用。

Objective： To observe the effects of Kangxian Ruangan granule on the hepatic stellate cell cycle of rat. Methods： SD male rats were gavaged Kangxian Ruangan granule for preparation of a medicament serum. HSC-T6 cell line placed in the incubator at 37℃under 5% CO2 in culture, were divided into drug group （ containing 5 mg/ml, 2. 5mg/ml, 1.25mg/ml three coflcentration of Kangxian Ruangan granule） , drug serum group （plus different concentrations Kangxian Ruan- gan granule drug serum, 5% , 10% , 20% ） , blank control group （plus DMEM medium containing 5% FCS） and normal serum （ plus normal rat serum） . The inhibition of cell proliferation rate was measured by MTY assay and the cell cycle was measured by flow cytometry. Results： The activation of HSC proliferation were inhibited both in the drug group and serum containing, the drug group and the drug-containing serum group were significantly better than the control group （ P 〈 0. 05, P 〈 0.01 ） . Cells in GO/G1 phase DNA content in comparison, 10% and 20% of the drug-containing serum group were significantly higher than the normal serum group （P 〈0. 05, P 〈 0.01 ）, the drug group were significantly higher than the control group （P 〈 0. 05, P 〈 0. 01 ） . Conclusion： Kangxian Ruangan granule can block HSC conducted from G1 to S phase （ block HSC through the G1/S checkpoint） , resulting in the accumulation of cells in G1 phase block DNA synthesis and replication, play a role in inhibiting the proliferation of HSC.